Mail: NW 311.13
52 Oxford St
Cambridge, MA 02138
Gaudet Lab Homepage
Members of the Gaudet Lab
List of Publications from PubMed
Term: Spring Term 2013-2014. Credit: Half course.
Instructors: Rachelle Gaudet, Andres Leschziner
Course Level: Primarily for Undergraduates
Description: The course aims to develop fundamental concepts of biochemistry as they apply to macromolecules, including protein and nucleic acid structure, thermodynamics and kinetics, ligand interactions and chemical equilibria. The course will also emphasize how these concepts are used in studies of the structure and function of biological molecules, including examples from metabolism. In the weekly section, students will undertake a discovery-based laboratory research project in which they will apply these concepts toward understanding the structure and function of the ATPase domain from the ABC transporter associated with antigen processing (TAP).
Prerequisite(s): LPSA or LS1a, Chemistry 20/30 or Chemistry 17/27 (Chemistry 27 may be concurrent), Math 1b. A solid foundation in molecular and cellular biology (which could be provided by MCB 52 or MCB 54, for example) and/or physics (e.g. PS2) is recommended.
Meetings: M., W., F., at 10, and a weekly laboratory/discussion section.
Term: Fall Term 2013-2014. Credit: Half course.
Instructors: Rachelle Gaudet, Andres Leschziner
Course Level: Primarily for Graduates
Description: This course will introduce basic principles in general, organic and physical chemistry, including kinetics and thermodynamics, as well as macromolecular structure. Concepts will be illustrated with examples taken from the visual system.
Note: Required for first year graduate students in the Molecules, Cells and Organisms (MCO) Training Program.
Meetings: M., W., 2-4
Term: Fall Term; Repeated Spring Term 2013-2014. Credit: Half course.
Instructor: Rachelle Gaudet
Course Level: Graduate Course
Structural biology of ion channels and transporters
We use a combination of x-ray crystallography and other biophysical and biochemical techniques to study the stereochemistry of signaling and transport through biological membranes.
Structural studies of TRP channels
The goal is to elucidate the gating mechanism of TRP ion channels involved in temperature sensing and understand modulatory interactions of proteins and small molecules with TRP channels. We are particularly interested in determining the molecular mechanism of temperature sensing. We therefore focus on the temperature-sensing TRP channels such as TRPV1, TRPV2, and TRPM8. Several temperature-sensing TRP channels like TRPV1, TRPV2 and TRPA1 are expressed in nociceptor neurons, and therefore responsible for pain sensations in response to noxious stimuli. The biophysical and biochemical mechanisms of pain and heat sensing are not only of academic interest, but also of medical and pharmacological interest. With structures of these TRP channels, we will be better equipped to design chemical inhibitors, which could potentially be used therapeutically.
Structural studies of TAP, the ABC transporter associated with antigen processing
The goal is to elucidate how TAP, a heterodimer of two membrane-spanning proteins, TAP1 and TAP2, transports peptides generated by the proteasome in the cytosol into the endoplasmic reticulum for loading onto MHC class I molecules. Loaded class I molecules then travel to the cell surface and present the peptides to T cells, an immune system mechanism to recognize and eliminate deregulated or tumorigenic cells, virally-infected cells and foreign cells (e.g. graft rejection). We have undertaken structural studies of the cytosolic C-terminal nucleotide bingin domains (NBDs) of TAP1 and TAP2 to understand how ATP binding and hydrolysis fuels peptide transport. Furthermore, we have overexpressed the full-length TAP ABC transporter at levels adequate for biochemical and crystallographic studies. We also use cell-based and in vitro biochemical assays to determine the effects of deletions and substitutions within the TAP1 and TAP2 proteins.
Structural studies of Nramp proteins
Metals such as iron and manganese are essential to physiological processes such as oxygen transport and energy metabolism. Nramps (natural resistance-associated macrophage proteins) are transporters that allow the proton-driven import of divalent metal ions into cells. Humans have two Nramp homologs. Nramp1 transports metals across the phagolysosomal membrane of macrophages and other phagocytic cells, and is important for the antimicrobial function of these cells. DMT1 (divalent metal transporter 1, also known as Nramp2 or DCT1) is responsible for absorption of dietary iron and manganese in the proximal duodenum. Furthermore, DMT1 allows assimilation of transferrin-bound iron by the red blood cell precursors in specialized endosomal compartments. The goal is to determine the molecular mechanism of metal-ion transport by the Nramp family of proteins through structural studies of a bacterial Nramp protein, MntH.
E. Procko, R. Gaudet (2009) Antigen processing and presentation: TAPping into ABC transporters. Curr Opin Immunol. 21, 84-91.
R. Gaudet (2009) Divide and conquer: high resolution structural information on TRP channel fragments. J Gen Physiol. 133, 231-7.
E. Procko, M. L. O’Mara, W. F. D. Bennett, D. P. Tieleman, R. Gaudet (2009) The mechanism of ABC transporters: general lessons from structural and functional studies of an antigenic peptide transporter. FASEB Journal 23, 1287-302.
E. Procko and R. Gaudet (2008) Functionally Important Interactions between the Nucleotide-Binding Domains of an Antigenic Peptide Transporter. Biochemistry 47, 5699-708.
C. B. Phelps, R. J. Huang, P. V. Lishko, R. R. Wang and R. Gaudet (2008) Structural analyses of the Ankyrin Repeat Domain of TRPV6 and related TRPV ion channels. Biochemistry 47, 2476-2484.
P. V. Lishko, E. Procko, X. Jin, C. B. Phelps and R. Gaudet (2007) The Ankyrin Repeats of TRPV1 Bind Multiple Ligands and Modulate Channel Sensitivity. Neuron 54, 905-918.
C. Schlieker, W. A. Weihofen, E. Frijns, L. M. Kattenhorn, R. Gaudet and H. L. Ploegh (2007) Structure of a Herpesvirus-Encoded Cysteine Protease Reveals a Unique Class of Deubiquitinating Enzymes. Mol. Cell 25, 677-687.
E. Procko, I. Ferrin-O’Connell, S.-L. Ng, and R. Gaudet (2006) Distinct structural and functional properties of the ATPase sites in an asymmetric ABC transporter. Mol. Cell 24, 51-62.
X. Jin, J. Touhey, and R. Gaudet (2006) Structure of the N-terminal ankyrin repeat domain of the TRPV2 ion channel. J. Biol. Chem. 281, 25006-25010.